Bodo sp., a Free‐Living Flagellate, Expresses Divergent Proteolytic Activities from the Closely Related Parasitic Trypanosomatids

Abstract

We report the characterization of cell-associated and extracellular peptidases of Bodo sp., a free-living flagellate of the Bodonidae family, order Kinetoplastida, which is considered ancestral to the trypanosomatids. This bodonid isolate is phylogenetically related to Bodo caudatus and Bodo curvifilus. The proteolytic activity profiles of Bodo sp. were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing co-polymerized gelatin, casein, hemoglobin, or bovine serum albumin as substrates. The enzymatic complex degraded gelatin better in acidic pH, and under these conditions four proteolytic bands (120, 100, 90, and 75 kDa) were detected in the cellular or extracellular extracts. Two peptidases (250 and 200 kDa) were exclusively detected with the substrate casein. All these enzymes belong to the serine peptidase class, based on inhibition by aprotinin and phenylmethylsulfonyl fluoride. This is the first biochemical characterization of peptidases in a free-living Bodo sp., potentially providing insight into the physiology of these protozoa and the evolutionary importance of peptidases to the order Kinetoplastida as some of these enzymes are important virulence factors in pathogenic trypanosomatids.

RNA-Seq-Based Metatranscriptomic and Microscopic Investigation Reveals Novel Metalloproteases of Neobodo sp. as Potential Virulence Factors for Soft Tunic Syndrome in Halocynthia roretzi

“…However, in the Kinetoplastida, most studies have focused on trypanosomatids [1], [3]. Only one group previously characterized the proteases of a Bodo sp., and their results suggested potential differences in protease expression among the kinetoplastida [73].…”

Genes codificadores de proteínas implicadas na relação de espécies do gênero <i>Trypanosoma</i> com seus hospedeiros: diversidade, transferência horizontal e relações filogenéticas.

“…Os bodonídeos são em geral organismos biflagelados e de vida livre, exceto os dos gêneros Cryptobia e Trypanoplasma, que são parasitas de peixes. As espécies de vida livre são encontradas tanto no solo quanto em ambientes aquáticos, de água doce e salgada, e atuam como micropredadores que auxiliam no equilíbrio microbiológico destes ambientes (D'Avila-Levy et al, 2009). Os bodonídeos são divididos em vários gêneros, entre eles: Perkinsiella, Bodo, Neobodo, Parabodo, Trypanoplasma, Dimastigella, Rhynchobodo, Rhynchomonas e Cryptobia (López-García et al, 2003;Moreira et al, 2004;von der Heyden, Cavalier-Smith, 2005).…”